In this research, by using aptamer-conjugated gold nanoparticles (aptamer-AuNPs) and a modified glassy carbon electrode (GCE) with reduced graphene oxide (rGO) and Acropora-like gold (ALG) nanostructure, a sandwich-like system provided for sensitive detection of heat shock protein 70 kDa (HSP70), which applied as a functional biomarker in diagnosis/prognosis of COVID-19. Initially, the surface of the GCE was improved with rGO and ALG nanostructures, respectively. Then, an aptamer sequence as the first part of the bioreceptor was covalently bound on the surface of the GCE/rGO/ALG nanostructures. After adding the analyte, the second part of the bioreceptor (aptamer-AuNPs) was immobilized on the electrode surface to improve the diagnostic performance. The designed aptasensor detected HSP70 in a wide linear range, from 5 pg mL−1 to 75 ng mL−1, with a limit of detection (LOD) of ∼2 pg mL−1. The aptasensor was stable for 3 weeks and applicable in detecting 40 real plasma samples of COVID-19 patients. The diagnostic sensitivity and specificity were 90% and 85%, respectively, compared with the reverse transcription-polymerase chain reaction (RT-PCR) method. •First biosensing application of HSP70 for diagnosis/prognosis of COVID-19 developed.•Sandwich-like design equipped with reduced graphene oxide (rGO), Acropora-like gold (ALG) nanostructure, and aptamer-AuNPs.•Aptasensor confirmed a wide linear detection range (5 pg mL-1 - 75 ng mL-1) and a desirable LOD (2 pg mL-1).•Forty real plasma samples of hospitalized COVID-19 patients were analyzed and compared to the RT-PCR method.